Abstract
Tuberculosis (TB) is an infectious disease in which the molecular typing methods allow to have important information about the dynamics of transmission and to assist properly in disease control. Although the ERIC-PCR (Enterobacterial repetitive intergenic consensus-PCR) assay is fast and easy to perform, scarce studies have reported its use in epidemiological studies in TB outbreaks. In this study, we aimed to genotype Mycobacterium tuberculosis and M. bovis isolates by ERIC-PCR and compare its discriminatory power with two other classically used methods: 12 loci-MIRU (Mycobacterial Interspersed Repetitive Units) and Spoligotyping. The M. tuberculosis isolates studied were from northwestern and southwestern and M. bovis from northwestern Parana, Brazil. ERIC-PCR rendered banding patterns with great diversity (1 to 12 bands) of molecular sizes, ranging from 100 to 1600 bp. ERIC-PCR showed to be fast, simple and affordable to differentiate isolates. ERIC-PCR would be an important tool in the epidemiology of TB as screening in case of outbreak, which demands rapid intervention. However if any doubt persist, as it may occur with the application of only one genotypic method, other genotyping methods should be applied and carefully interpreted, always with additional epidemiological information.
Highlights
Tuberculosis (TB) is an infectious disease caused mainly by Mycobacterium tuberculosis and it has been known for centuries
The application of Spoligotyping plus Enterobacterial Repetitive Intergenic Consensus sequence (ERIC)-PCR in 73 isolates could not differentiate only 2 clustered isolates The analysis of 63 isolates genotyped by 12 loci-MIRU plus ERIC-PCR showed 100 % of differentiation (Figure 2)
A very simple and fast genotyping method, ERICPCR, with Spoligotyping and MIRU to differentiate epidemiologically related from unrelated M. tuberculosis isolates
Summary
Tuberculosis (TB) is an infectious disease caused mainly by Mycobacterium tuberculosis and it has been known for centuries. Molecular differentiation of isolates is useful in epidemiological investigations as it is a means to better understand the mechanisms that influence the dynamics of transmission, identification of risk factors in a specific community and assist properly in disease control. The genetic differentiation of M. tuberculosis complex isolates can determine the source of outbreaks and the relation between TB in domestic and wild animals, and identify the source of human infections (Mears et al, 2015). A highly discriminatory typing method based on variations of the IS6110 specific sequences in the M. tuberculosis genome, the Restriction Fragment Lenght Polymorfism (RFLP) has been widely used in epidemiological studies of TB. Despite the promising results, the RFLP is laborious, requiring highly skilled personnel to carry out the trial
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